Effects of Active Extracts of Miaoyao Fanggan Sachet on bronchial epithelial cells stimulated with polymyocytic acid

doi:10.3877/cma.j.issn.1674-0785.2021.04.007

Abstract

Abstract:

Objective

To investigate the effects of diammonium glycyrrhizinate (DG) and magnesium isoglycyrrhizinate (MI) on bronchial epithelial cells (NHBE) stimultated with polymyocytic acid (PIC), to determine the therapeutic effects of DG and MI and to explore the possible mechanisms involved.

Methods

NHBE cells were stimulated with 1 μg/ml PIC to construct the cell model. Cells treated with Dexamethasone (DXMS) at 10 μg/ml were used as a positive control. DG and MI at low, medium, and high concentrations (0.5, 1.0, 2.0 mg/ml) were used for cell intervention. Cell activity was detected with CCK8 kit. IFN-α and IL-6 levels were detected by ELISA. IFN-α and IL-6 mRNA levels were detected by qPCR. p-TAK1/TAK1 and p-IkB/IkB expression levels were detected by Western blot.

Results

CCK8 assay showed that the viability of NHBE cells decreased with the increase of drug concentration, and the decrease of cell viability was more obvious in the positive control group. Compared with blank control cells, the expression of IFN-α and IL-6 significantly increased in the model group (cells treated with PIC; P＜0.05). Compared with the model group, the expression levels of IFN-α and IL-6 significantly decreased after treatment with different doses of DG and MI (P＜0.05). The mRNA expression levels of IFN-α and IL-6 in the model group were significantly higher compared with the blank control group (P＜0.05), but these in the intervention groups were significantly decreased (P＜0.05). After modeling, the ratios of p-TAK1/TAK1 and p-IkB/IkB tended to increase significantly (P＜0.05), while they showed no signficant changes in NHBE cells treated with low, medium, and high concentrations of DG and MI. In the positive control group, these ratios decreased compared with those in the model group (P＜0.05).

Conclusion

Ammonium glycyrrhizinate is one of the main active components of Miaoyao Fanggan Sachet. DG and MI have certain curative effects on PIC induced NHBE proinflammatory response possibly by downregulating IFN-α and IL-6 expression and reducing the inflammatory response induced by PIC. Whether the mechamism of PIC induced pro-inflammatory response of NHBE cells is related to the phosphorylation levels of TAK1 and IKB proteins needs to be further studied.

Key words: Miaoyao Fanggan Sachet, Polymyocytic acid, Diammonium glycyrrhizinate, Magnesium isoglycyrrhizinate

Xiaofen Zhang, Quan Zhang. Effects of Active Extracts of Miaoyao Fanggan Sachet on bronchial epithelial cells stimulated with polymyocytic acid[J]. Chinese Journal of Clinicians(Electronic Edition), 2021, 15(04): 272-279.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhlcyszz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-0785.2021.04.007

https://zhlcyszz.cma-cmc.com.cn/EN/Y2021/V15/I04/272

Figures/Tables 9

References 16

1	张景熙, 焦洋, 宁允叶, 等. 地塞米松抑制聚肌胞苷酸诱导人气道上皮细胞趋化因子表达及机制的初步研究 [J]. 第二军医大学学报, 2012, 33(8): 824-828.
2	Swindle EJ, Collins JE, Davies DE. Breakdown in epithelial barrier function in patients with asthma: identification of novel therapeutic approaches [J]. J Allergy Clin Immunol, 2009, 124(1): 23-34.
3	Torres D, Dieudonné A, Ryffel B, et al. Double-stranded RNA exacerbates pulmonary allergic reaction through TLR3: implication of airway epithelium and dendritic cells [J]. J Immunol, 2010, 185(1): 451-459.
4	Eddleston J, Lee RU, Doerner AM, et al. Cigarette smoke decreases innate responses of epithelial cells to rhinovirus infection [J]. Am J Respir Cell Mol Biol, 2011, 44(1): 118-126.
5	Choi JP, Kim YS, Tae YM, et al. A viral PAMP double-stranded RNA induces allergen-specific Th17 cell response in the airways which is dependent on VEGF and IL-6 [J]. Allergy, 2010, 65: 1322-1330.
6	金鸣昌, 晏志, 郭伟伟, 等. 苗药防感香囊预防甲型H1N1流感的应用观察 [J]. 中国中医药信息杂志, 2013, 20(6): 63-64.
7	韩飞, 彭珍, 周志渝, 等. 功效性分类中药对提高机体免疫功能的研究进展 [J]. 中草药, 2016, 47(14): 2549-2555.
8	Walther B, Sieber R. Bioactive proteins and peptides in foods [J]. Int J Vitam Nutr Res, 2011, 81(2/3): 181-192.
9	Pestka S, Krause CD, Walter MR. Interferons, interferon-like cytokines, and their receptors [J]. Immunol Rev, 2004, 202: 8-32.
10	Lamken P, Gavutis M, Peters I, et al. Functional cartography of the ectodomain of the type I interferon receptor subunitifnar1 [J]. J Mol Biol, 2005, 350(3):476-488.
11	Garbers C, Scheller J. Interleukin-6 and interleukin-11: same but different [J]. Biol Chem, 2013, 394(9): 1145-1161.
12	Wolf J, Waetzig GH, Reinheimer TM, et al. A soluble form of the interleukin-6 family signal transducer gp130 is dimerized via a C-terminal disulfide bridge resulting from alternative mRNA splicing [J]. Biochem Biophys Res Commun, 2016, 470(4): 870-876.
13	Pantano C, Ather JL, Alcorn JF, et al. Nuclear factor-kappaB activation in airway epithelium induces inflammation and hyperresponsiveness [J]. Am J Respir Crit Care Med, 2008, 177(9): 959-969.
14	Ali S, Hirschfeld AF, Mayer ML, et al. Functional genetic variation in NFKBIA and susceptibility to childhood asthma, bronchiolitis, and bronchopulmonary dysplasia [J]. J Immunol, 2013, 190(8): 3949-3958.
15	Ismail HM, Didangelos A, Vincent TL, et al. Articular cartilage injury rapidly activates TAK1 in chondrocytes by inducing its phosphorylation and K63-polyubiquitination [J]. Arthritis Rheumatol, 2017, 69(3): 565-575.
16	Mao RF, Fan YH, Mou YG. TAK1 lysine 158 is required for TGF-β-induced TRAF6-mediated Smad-independent IKK/NF-κB and JNK/AP-1 activation [J]. Cell Signal, 2011, 23(1): 222-227.

引物名称	引物序列	引物长度（bp）	产物长度（bp）	退火温度（℃）
IL-6F	CTTCGGTCCAGTTGCCTTCT	20	228	59.65
IL-6R	GCCTCTTTGCTGCTTTCACA	20	228	59.65
IFN-αF	GAAACTCCCCTGATGAATGC	20	119	57.55
IFN-αR	TCTGCTCTGACAACCTCCC	19	119	57.55
β-actinF	TGGCACCCAGCACAATGAA	19	186	60.8
β-actinR	CTAAGTCATAGTCCGCCTAGAAGCA	25	186	60.8

引物名称	引物序列	引物长度（bp）	产物长度（bp）	退火温度（℃）
IL-6F	CTTCGGTCCAGTTGCCTTCT	20	228	59.65
IL-6R	GCCTCTTTGCTGCTTTCACA	20	228	59.65
IFN-αF	GAAACTCCCCTGATGAATGC	20	119	57.55
IFN-αR	TCTGCTCTGACAACCTCCC	19	119	57.55
β-actinF	TGGCACCCAGCACAATGAA	19	186	60.8
β-actinR	CTAAGTCATAGTCCGCCTAGAAGCA	25	186	60.8

组分名称	质量含量（mg/kg）	俗称（CAS No.）
癸酸	1.5×10^-4	334-48-5
甘草酸铵盐	0.14~0.16	1405-86-3
蛋白质	0.05	-
总糖	0.19	-
蒎烯	微量	7785-70-8
β-蒎烯	微量	8172-67-3
P-伞花烃	微量	535-77-3
桉叶油醇	微量	470-82-6
左旋樟脑	微量	464-48-2
L-薄荷醇	微量	2216-51-5
α-松油醇	微量	10492-56-1
丙氨酸	9.69	-
钾元素	46.7	-
铜元素	4.4	-
钙元素	7.7	-

组分名称	质量含量（mg/kg）	俗称（CAS No.）
癸酸	1.5×10^-4	334-48-5
甘草酸铵盐	0.14~0.16	1405-86-3
蛋白质	0.05	-
总糖	0.19	-
蒎烯	微量	7785-70-8
β-蒎烯	微量	8172-67-3
P-伞花烃	微量	535-77-3
桉叶油醇	微量	470-82-6
左旋樟脑	微量	464-48-2
L-薄荷醇	微量	2216-51-5
α-松油醇	微量	10492-56-1
丙氨酸	9.69	-
钾元素	46.7	-
铜元素	4.4	-
钙元素	7.7	-

[1]	Da Wu, Zhongquan Tang. Clinical observation of magnesium isoglycyrrhizinate prevention on liver damage caused by Brucellosis [J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2016, 10(05): 599-602.
[2]	Xueying Lai, Bin Liu, Xueqin Hu, Haojun Chen. Therapeutic effect of magnesium isoglycyrrhizinate on liver injury induced by paclitaxel and its effect on serum IL-6, IL-10 and TNF-α [J]. Chinese Journal of Clinicians(Electronic Edition), 2020, 14(11): 922-925.
[3]	Huigang Feng, Qiwei Liang, Huizhuang Guo, Weizhao Zhuang, Chen Huang, Ying Tang, Chaoxiong He, Yi Huang. Clinical observation on diammonium glycyrrhizinate combined with huaier granule in the treatment of embolism syndrome after TACE therapy on hepatocellular carcinoma [J]. Chinese Journal of Interventional Radiology(Electronic Edition), 2016, 04(04): 197-201.

Please choose a citation manager

Content to export